The vitelline membrane or vitelline envelope is a structure surrounding the outer surface of the plasma membrane of an ovum (the oolemma) or, in some animals (e.g., birds), the extracellular yolk and the oolemma. It is composed mostly of protein fibers, with protein receptors needed for sperm binding which, in turn, are bound to sperm plasma membrane receptors. The species-specificity between these receptors contributes to prevention of breeding between different species. It is called zona pellucida in mammals. Between the vitelline membrane and zona pellucida is a fluid-filled perivitelline space.

As soon as the spermatozoon fuses with the ovum, signal transduction occurs, resulting in an increase of cytoplasmic calcium ions. This itself triggers the cortical reaction, which results in depositing several substances onto the vitelline membrane through exocytosis of the cortical granules, transforming it into a hard layer called the “fertilization membrane”, which serves as a barrier inaccessible to other spermatozoa. This phenomenon is the slow block to polyspermy.

In insects, the vitelline membrane is called the vitelline envelope and is the inner lining of the chorion.